Aim To develop and validate a liquid chromatographic-tandem mass spectrometric（LC-MS/MS） method for the determination of risperidone in human plasma.Methods Risperidone and the internal standard,diphenhydramine, wer...Aim To develop and validate a liquid chromatographic-tandem mass spectrometric（LC-MS/MS） method for the determination of risperidone in human plasma.Methods Risperidone and the internal standard,diphenhydramine, were isolated from plasma by liquid-liquid extraction with ether-dichloromethane（3∶2,v/v）,then chromatographed on a Zorbax Extend-C18 column（150 mm×4.6 mm ID,5 μm） using a mobile phase consisted of acetonitrile-water-formic acid（40∶60∶0.5,v/v）,at a flow rate of 0.7 mL·min-1.A Finnigan TSQ tandem mass spectrometer equipped with atmospheric pressure chemical ionization source was used as detector and was operated in the positive ion mode.Selected reaction monitoring（SRM） using the precursor product ion combinations of m/z 411→191 and m/z 256→167 were used to quantify risperidone and diphenhydramine（IS）,respectively.Results The linear concentration range of the calibration curve for risperidone was 0.025-50 μg·L-1.The lower limit of quantification was 0.025 μg·L-1.The intra-and inter-day relative standard deviation（RSD） across three validation running over the entire concentration range was less than 7.1%.The accuracy was within（±3.8%）.Each sample was chromatographed within 2.7 min.Conclusion The method was proved to be rapid,sensitive and suitable for pharmacokinetic investigations of risperidone.展开更多
Aim To develop and validate a liquid chromatography-tandem mass spectrometric（LC/MS/MS） method for the simultaneous quantification of ephedrine and chlorpheniramine in human plasma after oral administration of a com...Aim To develop and validate a liquid chromatography-tandem mass spectrometric（LC/MS/MS） method for the simultaneous quantification of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation.Methods The analytes and the internal standard,diphenhydramine,were isolated from plasma by protein precipitation with methanol,then chromatographied on a Zorbax SB-C18 column（150 mm×4.6 mm ID） using a mobile phase consisted of methanol-water-formic acid（80∶20∶0.5,v/v）,at a flow rate of 0.5 mL·min-1.A tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode.Selected reaction monitoring（SRM） using the precursor to produce ion combinations of m/z 166→115,（m/z） 275→230 and m/z 256→167 were used to quantify ephedrine,chlorpheniramine and the internal standard,respectively.Results The linear concentration ranges of the calibration curves for ephedrine and chlorpheniramine were 0.50-200 μg·L-1 and 0.050-20.0 μg·L-1,respectively.The lower limits of quantification were 0.50 μg·L-1 for ephedrine and 0.050 μg·L-1 for chlorpheniramine,individually.The intra-and inter-day relative standard deviation（RSD） across three validation runs over the entire concentration range was less than 9.3% for both ephedrine and chlorpheniramine.The inter-day accuracy（RE） was within ±3.4% for the analytes.Each sample was chromatographied within 3.3 min.The method was successfully used in pharmacokinetics study of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation containing 5 mg ephedrine hydrochloride,1 mg chlorpheniramine maleate,50 mg phenytoin,12.5 mg theophylline,12.5 mg theobromine and 7.5 mg caffeine.No interaction among the six components was observed on their pharmacokinetic parameters.Conclusion The method was proved to be highly sensitive,selective,and suitable for pharmacokinetics investigations of different compound preparations containing low dosage of both ephedrine and chlorpheniramine.展开更多
目的揭示肥胖诱发脂肪组织线粒体蛋白质组的变化,并探索变化的可能机制,实现针对小鼠脂肪组织线粒体蛋白质组的深度测序和准确定量。方法利用数据依赖型(DDA)质谱方法鉴定小鼠白色脂肪组织和棕色脂肪组织中的线粒体蛋白;利用新型SWATH(s...目的揭示肥胖诱发脂肪组织线粒体蛋白质组的变化,并探索变化的可能机制,实现针对小鼠脂肪组织线粒体蛋白质组的深度测序和准确定量。方法利用数据依赖型(DDA)质谱方法鉴定小鼠白色脂肪组织和棕色脂肪组织中的线粒体蛋白;利用新型SWATH(sequential windowed acquisition of all theoretical mass spectra)质谱技术定量肥胖小鼠和野生型小鼠不同脂肪组织中线粒体蛋白,同时对表达差异的线粒体蛋白进行功能分析。实验步骤:1用组织线粒体提取试剂盒特异性提取不同类型脂肪组织线粒体蛋白;2用10%聚丙烯酰胺凝胶电泳预分离线粒体蛋白;3利用DDA方法建立线粒体蛋白质谱文库;4利用SWATH质谱方法定量线粒体蛋白;5利用Peak View2.0软件提取碎片离子色谱峰并进行积分计算其峰面积;6根据KEGG(kyoto encyclopedia of genes and genomes)通路和GO(Gene Ontology)功能分类分析肥胖小鼠和正常小鼠脂肪组织表达差异蛋白的生物功能。结果在白色脂肪组织和棕色脂肪组织分别准确鉴定并定量线粒体定位蛋白1000和1039种,3次生物学重复实验中共鉴定包括线粒体氨肟还原成分1在内的25种线粒体蛋白在白色脂肪组织特异性表达,包括非偶联蛋白3在内的21种线粒体蛋白在棕色脂肪组织特异性表达。肥胖小鼠与正常小鼠相比,白色脂肪组织中共有25种线粒体蛋白表达显著上调(上调倍数≥2,P<0.01),有47种线粒体蛋白表达显著下调(下调倍数≥2,P<0.01);棕色脂肪组织中有26种线粒体蛋白表达显著上调(上调倍数≥2,P<0.01),有21种线粒体蛋白表达显著下调(下调倍数≥2,P<0.01)。结论脂肪组织线粒体蛋白质组的定量及生物信息学的分析表明,肥胖导致线粒体内三羧酸循环、氧化磷酸化、脂肪酸的合成和降解以及对外源性物质的代谢等重要的信号通路发生紊乱,并准确鉴定、定量其相关表达差异蛋白,为肥胖导致代谢紊乱的机制研究提供重要的数据支持。展开更多
探究9次跨膜超家族蛋白2(transmembrane 9 superfamily protein member 2,TM9SF2)对于三阴性乳腺癌MDA-MB-231细胞增殖和转移的影响及其分子机制。采用Western blot实验检测三阴性乳腺癌细胞株MDA-MB-231和非致瘤的乳腺上皮细胞株MCF-10...探究9次跨膜超家族蛋白2(transmembrane 9 superfamily protein member 2,TM9SF2)对于三阴性乳腺癌MDA-MB-231细胞增殖和转移的影响及其分子机制。采用Western blot实验检测三阴性乳腺癌细胞株MDA-MB-231和非致瘤的乳腺上皮细胞株MCF-10A中TM9SF2蛋白表达的情况;对高表达TM9SF2的三阴性细胞株MDA-MB-231进行基因沉默;采用MTS法检测细胞增殖活性,采用Transwell实验和划痕实验检测细胞的转移能力;采用Western blot实验检测细胞内增殖相关蛋白(PI3K、AKT、SRC和ERK)和转移相关蛋白(Snail、Slug和N-cadherin)的表达情况。Western blot实验证明,MDA-MB-231中TM9SF2蛋白的表达量高于MCF-10A细胞。与对照组相比,siRNA-TM9SF2转染组TM9SF2蛋白表达下调,细胞增殖活性降低,细胞转移能力减弱,PI3K、Snail、Slug和N-cadherin表达水平均降低,AKT蛋白磷酸化激活降低。研究结果表明,TM9SF2基因能促进三阴型乳腺癌MDA-MB-231细胞的增殖和转移。展开更多
文摘Aim To develop and validate a liquid chromatographic-tandem mass spectrometric（LC-MS/MS） method for the determination of risperidone in human plasma.Methods Risperidone and the internal standard,diphenhydramine, were isolated from plasma by liquid-liquid extraction with ether-dichloromethane（3∶2,v/v）,then chromatographed on a Zorbax Extend-C18 column（150 mm×4.6 mm ID,5 μm） using a mobile phase consisted of acetonitrile-water-formic acid（40∶60∶0.5,v/v）,at a flow rate of 0.7 mL·min-1.A Finnigan TSQ tandem mass spectrometer equipped with atmospheric pressure chemical ionization source was used as detector and was operated in the positive ion mode.Selected reaction monitoring（SRM） using the precursor product ion combinations of m/z 411→191 and m/z 256→167 were used to quantify risperidone and diphenhydramine（IS）,respectively.Results The linear concentration range of the calibration curve for risperidone was 0.025-50 μg·L-1.The lower limit of quantification was 0.025 μg·L-1.The intra-and inter-day relative standard deviation（RSD） across three validation running over the entire concentration range was less than 7.1%.The accuracy was within（±3.8%）.Each sample was chromatographed within 2.7 min.Conclusion The method was proved to be rapid,sensitive and suitable for pharmacokinetic investigations of risperidone.
文摘Aim To develop and validate a liquid chromatography-tandem mass spectrometric（LC/MS/MS） method for the simultaneous quantification of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation.Methods The analytes and the internal standard,diphenhydramine,were isolated from plasma by protein precipitation with methanol,then chromatographied on a Zorbax SB-C18 column（150 mm×4.6 mm ID） using a mobile phase consisted of methanol-water-formic acid（80∶20∶0.5,v/v）,at a flow rate of 0.5 mL·min-1.A tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode.Selected reaction monitoring（SRM） using the precursor to produce ion combinations of m/z 166→115,（m/z） 275→230 and m/z 256→167 were used to quantify ephedrine,chlorpheniramine and the internal standard,respectively.Results The linear concentration ranges of the calibration curves for ephedrine and chlorpheniramine were 0.50-200 μg·L-1 and 0.050-20.0 μg·L-1,respectively.The lower limits of quantification were 0.50 μg·L-1 for ephedrine and 0.050 μg·L-1 for chlorpheniramine,individually.The intra-and inter-day relative standard deviation（RSD） across three validation runs over the entire concentration range was less than 9.3% for both ephedrine and chlorpheniramine.The inter-day accuracy（RE） was within ±3.4% for the analytes.Each sample was chromatographied within 3.3 min.The method was successfully used in pharmacokinetics study of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation containing 5 mg ephedrine hydrochloride,1 mg chlorpheniramine maleate,50 mg phenytoin,12.5 mg theophylline,12.5 mg theobromine and 7.5 mg caffeine.No interaction among the six components was observed on their pharmacokinetic parameters.Conclusion The method was proved to be highly sensitive,selective,and suitable for pharmacokinetics investigations of different compound preparations containing low dosage of both ephedrine and chlorpheniramine.
文摘目的揭示肥胖诱发脂肪组织线粒体蛋白质组的变化,并探索变化的可能机制,实现针对小鼠脂肪组织线粒体蛋白质组的深度测序和准确定量。方法利用数据依赖型(DDA)质谱方法鉴定小鼠白色脂肪组织和棕色脂肪组织中的线粒体蛋白;利用新型SWATH(sequential windowed acquisition of all theoretical mass spectra)质谱技术定量肥胖小鼠和野生型小鼠不同脂肪组织中线粒体蛋白,同时对表达差异的线粒体蛋白进行功能分析。实验步骤:1用组织线粒体提取试剂盒特异性提取不同类型脂肪组织线粒体蛋白;2用10%聚丙烯酰胺凝胶电泳预分离线粒体蛋白;3利用DDA方法建立线粒体蛋白质谱文库;4利用SWATH质谱方法定量线粒体蛋白;5利用Peak View2.0软件提取碎片离子色谱峰并进行积分计算其峰面积;6根据KEGG(kyoto encyclopedia of genes and genomes)通路和GO(Gene Ontology)功能分类分析肥胖小鼠和正常小鼠脂肪组织表达差异蛋白的生物功能。结果在白色脂肪组织和棕色脂肪组织分别准确鉴定并定量线粒体定位蛋白1000和1039种,3次生物学重复实验中共鉴定包括线粒体氨肟还原成分1在内的25种线粒体蛋白在白色脂肪组织特异性表达,包括非偶联蛋白3在内的21种线粒体蛋白在棕色脂肪组织特异性表达。肥胖小鼠与正常小鼠相比,白色脂肪组织中共有25种线粒体蛋白表达显著上调(上调倍数≥2,P<0.01),有47种线粒体蛋白表达显著下调(下调倍数≥2,P<0.01);棕色脂肪组织中有26种线粒体蛋白表达显著上调(上调倍数≥2,P<0.01),有21种线粒体蛋白表达显著下调(下调倍数≥2,P<0.01)。结论脂肪组织线粒体蛋白质组的定量及生物信息学的分析表明,肥胖导致线粒体内三羧酸循环、氧化磷酸化、脂肪酸的合成和降解以及对外源性物质的代谢等重要的信号通路发生紊乱,并准确鉴定、定量其相关表达差异蛋白,为肥胖导致代谢紊乱的机制研究提供重要的数据支持。
文摘探究9次跨膜超家族蛋白2(transmembrane 9 superfamily protein member 2,TM9SF2)对于三阴性乳腺癌MDA-MB-231细胞增殖和转移的影响及其分子机制。采用Western blot实验检测三阴性乳腺癌细胞株MDA-MB-231和非致瘤的乳腺上皮细胞株MCF-10A中TM9SF2蛋白表达的情况;对高表达TM9SF2的三阴性细胞株MDA-MB-231进行基因沉默;采用MTS法检测细胞增殖活性,采用Transwell实验和划痕实验检测细胞的转移能力;采用Western blot实验检测细胞内增殖相关蛋白(PI3K、AKT、SRC和ERK)和转移相关蛋白(Snail、Slug和N-cadherin)的表达情况。Western blot实验证明,MDA-MB-231中TM9SF2蛋白的表达量高于MCF-10A细胞。与对照组相比,siRNA-TM9SF2转染组TM9SF2蛋白表达下调,细胞增殖活性降低,细胞转移能力减弱,PI3K、Snail、Slug和N-cadherin表达水平均降低,AKT蛋白磷酸化激活降低。研究结果表明,TM9SF2基因能促进三阴型乳腺癌MDA-MB-231细胞的增殖和转移。
